In vitro Transplantation of Fetal Human Retinal Pigment Epithelial Cells onto Human Cadaver Bruch's Membrane

Early Attachment of Uncultured Retinal Pigment Epithelium from Aged Donors onto Bruch's Membrane Explants

“…Previously it was found that cultured fetal RPE cells can repopulate human cadaver submacular Bruch's membrane explants debrided of native RPE (Castellarin et al, 1998b). These studies showed that fetal cells can attach rapidly and by 1 hr, with the same number of cells used in the present study, can effectively cover basement membrane surfaces (mean approximately 92 % coverage) and partially cover the inner collagenous layer surfaces (mean approximately 37 % coverage).…”

“…Within the range of cell density studied (approximately 120±480 Â 10 3 cells/7 mm diameter Bruch's membrane explant), seeding more cells increased the number of RPE attached only if the native RPE basement membrane was present. As the incubation time increased, however, the percentage of explant coverage by seeded RPE, while increasing signi®cantly in the presence of basement membrane, never approximated the approximately 100 % coverage shown in experiments using cultured fetal RPE cells 24 hr following seeding (Castellarin et al, 1998b). These results indicate that freshly isolated RPE cells from aged donors as harvested in this study, will not be able attach in suf®cient numbers to Bruch's membrane to repopulate a denuded surface even in the presence of basement membrane.…”

“…In vitro RPE attachment experiments using cultured human fetal RPE indicate that RPE adhesion to submacular Bruch's membrane lacking native RPE basement membrane is somewhat limited at early time points (Castellarin et al, 1998b). Del showed that the reattachment rate of cultured RPE from older donors is highest to the inner parts of peripheral Bruch's membrane and decreases as deeper layers are exposed.…”

“…with native RPE basement membrane present approximately 90 % of a Bruch's membrane explant is covered with RPE cells in 1 hr; with native RPE basement membrane absent: approximately 37 % of the explant surface is covered by 1 hr). Intact fetal RPE cells were observed on the inner collagenous layer as long as 48 hr after seeding (the longest time point observed in this study) (Castellarin et al, 1998b).…”

“…A pair of Bruch's membrane explants from the same donor as the RPE cells were prepared as described previously (Castellarin et al, 1998b). Anterior segments were removed following peripheral choroid±RPE complex isolation.…”